RESUMEN
Nonalcoholic fatty liver disease (NAFLD) is the hepatic manifestation of the metabolic syndrome and its prevalence increases continuously. Here, we reprogrammed fibroblasts of a high grade NAFLD patient with homozygous wildtype PNPLA3 genotype. The induced pluripotent stem cells (iPSCs) were characterized by immunocytochemistry, flow cytometry, embryoid body formation, pluritest, DNA-fingerprinting and karyotype analysis.
Asunto(s)
Células Madre Pluripotentes Inducidas/metabolismo , Lipasa/genética , Proteínas de la Membrana/genética , Enfermedad del Hígado Graso no Alcohólico/genética , Adulto , Genotipo , Homocigoto , Humanos , Masculino , Enfermedad del Hígado Graso no Alcohólico/patologíaRESUMEN
Considered a feature of the metabolic syndrome, nonalcoholic fatty liver disease (NAFLD), is associated with insulin resistance, type 2 diabetes, obesity and drug toxicity. Its prevalence is estimated at about 30% in western countries mainly due to sedentary life styles and high fat diets. Genome-wide association studies have identified polymorphisms in several genes, for example, PNPLA3, and TM6SF2 which confer susceptibility to NAFLD. Here, we review recent findings in the NAFLD field with a particular focus on published transcriptomics datasets which we subject to a meta-analysis. We reveal a common gene signature correlating with the progression of the disease from steatosis and steatohepatitis and reveal that lipogenic and cholesterol metabolic pathways are main actors in this signature. We propose the use of disease-in-a-dish models based on hepatocyte-like cells derived from patient-specific induced pluripotent stem cells (iPSC). These will enable investigations into the contribution of genetic background in the progression from NALFD to non-alcoholic steatohepatitis. Furthermore, an iPSC-based approach should aid in the elucidation of the function of new biomarkers, thus enabling better diagnostic tests and validation of potential drug targets. Stem Cells 2017;35:89-96.
Asunto(s)
Investigación Biomédica , Enfermedad del Hígado Graso no Alcohólico/patología , Animales , Progresión de la Enfermedad , Epigénesis Genética , Microbioma Gastrointestinal , Estudio de Asociación del Genoma Completo , HumanosRESUMEN
Primary fibroblasts from a healthy 19years old female were reprogrammed by transduction of retroviruses OCT4, SOX2, c-MYC and KLF4. iPSCs were characterized by immunocytochemistry, embryonic body-formation, DNA-fingerprint and karyotype analysis and comparative transcriptome analyses with the human embryonic stem cell line H1 revealed a Pearsons correlation coefficient of 0.8952.
Asunto(s)
Células Madre Pluripotentes Inducidas/citología , Diferenciación Celular , Línea Celular , Reprogramación Celular , Hibridación Genómica Comparativa , Dermatoglifia del ADN , Dermis/citología , Cuerpos Embrioides/citología , Cuerpos Embrioides/metabolismo , Femenino , Fibroblastos/citología , Vectores Genéticos/genética , Vectores Genéticos/metabolismo , Voluntarios Sanos , Células Madre Embrionarias Humanas/citología , Células Madre Embrionarias Humanas/metabolismo , Humanos , Células Madre Pluripotentes Inducidas/metabolismo , Cariotipo , Factor 4 Similar a Kruppel , Microscopía Fluorescente , Retroviridae/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Adulto JovenRESUMEN
Primary fibroblasts from a low grade steatosis patient were reprogrammed by transduction of a combination of two episomal-based plasmids OCT4,SOX2, c-MYC and KLF4. iPSCs were characterized by immunocytochemistry, embryonic body-formation, DNA-fingerprint karyotype analysis and comparative transcriptome analyses with the human embryonic stem cell line H1 revealed a Pearsons correlation of 0.9251.
Asunto(s)
Hígado Graso/patología , Células Madre Pluripotentes Inducidas/citología , Diferenciación Celular , Línea Celular , Reprogramación Celular , Dermis/citología , Cuerpos Embrioides/citología , Cuerpos Embrioides/metabolismo , Hígado Graso/genética , Femenino , Fibroblastos/citología , Humanos , Células Madre Pluripotentes Inducidas/metabolismo , Cariotipo , Factor 4 Similar a Kruppel , Microscopía Fluorescente , Análisis de Secuencia de ADN , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Primary fibroblasts from a high grade steatosis patient were reprogrammed by transduction of retroviruses OCT4, SOX2, c-MYC and KLF4. IPSCs were characterized by immunocytochemistry, embryoid body-formation, DNA-fingerprint, karyotype analysis and comparative transcriptome analyses with the human embryonic stem cell line H1 revealed a Pearsons correlation coefficient of 0.9287. Resource table.
Asunto(s)
Hígado Graso/patología , Células Madre Pluripotentes Inducidas/citología , Diferenciación Celular , Línea Celular , Reprogramación Celular , Dermatoglifia del ADN , Dermis/citología , Cuerpos Embrioides/citología , Cuerpos Embrioides/metabolismo , Hígado Graso/metabolismo , Fibroblastos/citología , Vectores Genéticos/genética , Vectores Genéticos/metabolismo , Humanos , Células Madre Pluripotentes Inducidas/metabolismo , Cariotipo , Factor 4 Similar a Kruppel , Masculino , Microscopía Fluorescente , Retroviridae/genética , Índice de Severidad de la Enfermedad , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Nonalcoholic fatty liver disease (NAFLD/steatosis) is a metabolic disease characterized by the incorporation of fat into hepatocytes. In this study, we developed an in vitro model for NAFLD based on hepatocyte-like cells (HLCs) differentiated from human pluripotent stem cells. We induced fat storage in these HLCs and detected major expression changes of metabolism-associated genes, as well as an overall reduction of liver-related microRNAs. We observed an upregulation of the lipid droplet coating protein Perilipin 2 (PLIN2), as well as of numerous genes of the peroxisome proliferator-activated receptor (PPAR) pathway, which constitutes a regulatory hub for metabolic processes. Interference with PLIN2 and PPARα resulted in major alterations in gene expression, especially affecting lipid, glucose, and purine metabolism. Our model recapitulates many metabolic changes that are characteristic for NAFLD. It permits the dissection of disease-promoting molecular pathways and allows us to investigate the influences of distinct genetic backgrounds on disease progression.